Team:Paris/Transduction overview strategy

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<span/ id="bottom">[https://2009.igem.org/ iGEM ] > [[Team:Paris#top | Paris]] > [[Team:Paris/Transduction_overview#top | Reception]] > [[Team:Paris/Transduction_overview_strategy#bottom | Our strategy]]
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<span/ id="bottom">[https://2009.igem.org/ iGEM ] > [[Team:Paris#top | Paris]] > [[Team:Paris/Transduction_overview#top | Receiving the message]] > [[Team:Paris/Transduction_overview_strategy#bottom | Our strategy]]
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== Overview  ==
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* [[Team:Paris/Transduction_overview#Overview#bottom | Introduction]]
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==Membran Fusion : Our Strategy==
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* [[Team:Paris/Transduction_overview_fusion#bottom |A. Fusion ]]
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** [[Team:Paris/Transduction_overview_fusion#A.1 Jun/Fos|A.1 jun/fos]]
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** [[Team:Paris/Transduction_overview_fusion#A.2 G3P|A.2 G3P]]
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** [[Team:Paris/Transduction_overview_fusion#A.3 Snares |A.3 Snares]]
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* [[Team:Paris/Transduction_overview_transduction#bottom |B. Transduction]]
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** [[Team:Paris/Transduction_overview_transduction#B.1 ABC transporters|B.1 ABC transporters]]
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** [[Team:Paris/Transduction_overview_transduction#B.2 Two-component system|B.2  Two Component System]]
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* [[Team:Paris/Transduction_overview_strategy#bottom |C. Our strategy]]
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* [[Team:Paris/Transduction_overview_construction#bottom |D. Construction]]
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===C. Our Strategy===
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<a class="menu_sub"href="https://2009.igem.org/Team:Paris/Transduction_overview#bottom"> Main </a>|
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<a class="menu_sub"href="https://2009.igem.org/Team:Paris/Transduction_overview_fusion#bottom"> Fusion</a>|
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<a class="menu_sub_active"href="https://2009.igem.org/Team:Paris/Transduction_overview_strategy#bottom"> Our strategy</a>|
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<a class="menu_sub"href="https://2009.igem.org/Team:Paris/Transduction_overview_construction#bottom"> Construction</a>
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====C.1 The Fec operon ====
 
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Few ABC transporter such as FecABCD (iron transporter) are able to induce a response regardless of the tranlocation, due to the activity of FecA, moreover some mutant can also have a constitutive expression of FecABCD .
 
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===Jun/Fos strategy===
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Our strategy is to fuse the mutated Jun (the one unable to form homodimer) to AIDA, the autotransporter (for the donnor cell). By this way, Jun will be export to the outer membrane and should be on vesicles.
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The second step is to make a fusion between Fos and AIDA (for the receiver bacteria). In this direction Fos will be trasport to the outer membrane .
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The plan would be to use FecA- mutant  receiver and FecA+ mutant donor to transfert the constitutive FecA protein to the receiver. In this case the receiver will express the FecABCD operon without being induce by ferric citrate in the medium , and so we could place under the control of the Fec ABCD promoter, which is called pfec, the gene sequence encoding for the response. For the moment a response that would be easy to detect is the fluorescence of the RFP and the biobrick BBa-J61002 is the perfect candidate to test the system.
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===g3p strategy===
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g3p is naturally exposed at the surface of the filamentous bacteriophage. The M13 phage has a high affinity for E.coli, and if we could place its g3p on the surface of the vesicles it could activate the fusion with the Outer membrane of the targeted bacteria.
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[[Image:Transduction_overview_fec.png|500px|center| fec operon induction]]
 
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Outer membrane protein A (OmpA) is a major structural protein of the outer membrane of Escherichia coli.
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'''problems :''' the message is unidirectionnal and unrepeatable. It would just be a proof of principle that a vesicle-mediated controlled communication is possible.
 
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====C.x The trick TCS====
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To conclude, our strategy is to fuse OmpA-Linker to g3p to export the g3p protein to the outer membrane in order to find them on the vesicles surface.
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We could ,with the help of Alfonso Jaramillo, design a synthetic PBP which could detect the substrate we choosed and activate a specific HK.
 
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The previous work done by Valencia in 2006 was to design vanilin-sensitive PBP and a network for a graduated response whereas we just need a proteic sensitive PBP and a binary type of response.
 
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Even if the idea of creating a synthethic protein was very attractive, it would have been difficult to have concrete result in just 3 months.
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Latest revision as of 22:41, 21 October 2009

iGEM > Paris > Receiving the message > Our strategy


Membran Fusion : Our Strategy


Jun/Fos strategy

Our strategy is to fuse the mutated Jun (the one unable to form homodimer) to AIDA, the autotransporter (for the donnor cell). By this way, Jun will be export to the outer membrane and should be on vesicles. The second step is to make a fusion between Fos and AIDA (for the receiver bacteria). In this direction Fos will be trasport to the outer membrane .

g3p strategy

g3p is naturally exposed at the surface of the filamentous bacteriophage. The M13 phage has a high affinity for E.coli, and if we could place its g3p on the surface of the vesicles it could activate the fusion with the Outer membrane of the targeted bacteria.


Outer membrane protein A (OmpA) is a major structural protein of the outer membrane of Escherichia coli.


To conclude, our strategy is to fuse OmpA-Linker to g3p to export the g3p protein to the outer membrane in order to find them on the vesicles surface.


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